Department of Chemistry , Wuhan University , Wuhan , Hubei 430072 , People's Republic of China.
State Key Laboratory of Hybrid Rice, Department of Cell and Developmental Biology, College of Life Sciences , Wuhan University , Wuhan , Hubei 430072 , People's Republic of China.
J Agric Food Chem. 2020 Feb 26;68(8):2588-2596. doi: 10.1021/acs.jafc.9b07623. Epub 2020 Feb 14.
Monosaccharides play important roles in plant growth and development, and their biofunctions are closely related to their endogenous contents. Therefore, the determination of monosaccharides is beneficial for the further study of monosaccharide biofunction. In this work, we developed a liquid chromatography-mass spectrometry analytical method assisted by a post-column derivatization technique (LC-PCD-MS) for the fast and automatic determination of 16 monosaccharides in samples. Post-column chemical derivatization of monosaccharides was performed by a reaction of monosaccharides with 4-benzylaminobenzeneboronic acid (4-PAMBA) through boronate ester formation in a three-way connector. 4-PAMBA worked as a derivatization reagent to improve the selectivity and sensitivity of monosaccharide detection by MS. The developed LC-PCD-MS method integrates LC separation, chemical derivatization, and MS detection in one run, thus greatly reducing the analysis time for each sample. The limits of detection and limits of quantification for 16 monosaccharides were in the range of 0.002-0.1 and 0.007-0.5 ng/mL, respectively. Good linearity was obtained from the linear regression, with a determination coefficient () ranging from 0.9928 to 1.0000. The relative recoveries were in the range of 80.7-117.8%, with the intra- and interday relative standard deviations less than 19.7 and 16.5%, respectively, indicating good accuracy and acceptable reproducibility of the method. Finally, the method was successfully applied to investigate the spatial and temporal distribution of 16 monosaccharides in the developing flower and germinating seed of .
单糖在植物生长发育中起着重要作用，其生物功能与其内源性含量密切相关。因此，单糖的测定有利于进一步研究单糖的生物功能。在这项工作中，我们开发了一种液相色谱-质谱联用分析方法（LC-PCD-MS），辅助柱后衍生技术（LC-PCD-MS），用于快速自动测定样品中的 16 种单糖。单糖的柱后化学衍生通过单糖与 4-苄基氨基苯硼酸（4-PAMBA）在三通连接器中的硼酸酯形成反应进行。4-PAMBA 作为衍生试剂，通过 MS 提高了单糖检测的选择性和灵敏度。所开发的 LC-PCD-MS 方法将 LC 分离、化学衍生和 MS 检测集成在一次运行中，从而大大缩短了每个样品的分析时间。16 种单糖的检出限和定量限范围分别为 0.002-0.1 和 0.007-0.5 ng/mL。线性回归得到了良好的线性关系，决定系数（r²）范围为 0.9928-1.0000。相对回收率在 80.7-117.8%范围内，日内和日间相对标准偏差分别小于 19.7%和 16.5%，表明该方法具有良好的准确性和可接受的重现性。最后，该方法成功应用于研究 16 种单糖在 发育花朵和发芽种子中的时空分布。
